Informace o publikaci

Characterisation of invisible conformation of domain 1.1 of σA factor of RNA polymerase from Bacillus subtilis

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Rok publikování 2023
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Popis Introduction: ? factors are essential components of bacterial RNA polymerase (RNAP) as they allow to recognize promotor sequences and initiate transcription. Domain 1.1 of vegetative ? factors occupies the primary channel of RNAP and also prevents binding of the ? factor to promoter DNA alone. Here, we show that domain 1.1 of Bacillus subtilis ?A exists in two structurally distinct variants in dynamic equilibrium. Aims: To elucidate the structure and dynamics of minor conformation and to discover how does minor conformation affect transcription. Methods: Relaxation dispersion analysis, chemical exchange saturation transfer analysis, in vitro transcription Results: The major conformation at room temperature is represented by a previously reported well-folded structure solved by nuclear magnetic resonance, but 4 % of the protein molecules are present in a less thermodynamically favorable state. We show that this population increases with temperature and we predict its significant elevation at higher but still biologically relevant temperatures. We found that, in contrast to the major state, the detected minor state is partially unfolded. Its propensity to form secondary structure elements is especially decreased for the first and third ? helices, while the second ? helix and ß strand close to the C-terminus are more stable. Functional experiments with full length ?A and its shortened version lacking domain 1.1(?A_?1.1) then revealed that while full length ?A increases transcription activity of RNAP with increasing temperature, transcription with ?A_?1.1 remains constant. Conclusions: In conclusion, this study reveals conformational dynamics of domain 1.1 and provides a basis for studies of its interaction with RNAP and effects on transcription regulation.
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